Strusovskaya O. G., Baykin P. I.

Northen State Medical University, Arkhangelsk

Chronic toxity determination of Cochlearia officinalis infusion

Actual task of modern pharmacy is creation of drugs on vegetable base, having a low threshold of undesirable effects beginning. Cochlearia officinalis is a plant, which potentially has a hypoglycemic activity. Plant extract’s hypoglycemic activity connected with availability in its chemical composition of calystegines – polyhydroxylated unesterified nortropan alkaloids, which are able to competitively inhibit activity of glucosidases [1]. C. officinalis chronic toxity studying is necessary because of long drug use necessity, influencing on postprandial conditions.

«Wistar» line adult rats weighing 180-200 grams were held in quarantine for 14 days before experiment and used for toxicological research. Experimental animals maintenance was corresponded to «Health rules on the device, equipment and maintenance of experimental biological vivariums» and to order of The Russian Federation Ministry of Health №267 of 19.06.2003 «Laboratory practicies rules adoption» [2,3]. Animals were kept with continuous access to water on a standard diet. Feeding was carried out in fixed time. Experiment was conducted according to guidance for preclinical research of new drugs [4]. C. officinalis infusion was prepared in accordance with requirement of State Pharmacopoeia XI [5].

The toxic doses of C. officinalis infusion six animals in each of four experimental and control groups were used in this study. Determination of chronic toxity was conducted by giving infusion to animals of experimental groups during 60 days. Parallely groups of intact animals were given 0,9 % solution of sodium chloridi in the same quantity. Observation of the animals was carried out daily. Basic fixed parameters were: animal weight, general state of health, frequency of eating, defecation and urination.

Dynamics of animals’ body mass change presents in table 1.

Thus, these studies are the base of conclusion, that using of C. officinalis

Table 1

Dynamics of animal’s body mass change

Days of experiment	Average mass of animals in control groups, grams		Average mass of animals in experimental groups, grams
	Female	Male	Female	Male
1	197,1±3,5	200,2±1,4	197,6±3,5	200,2±5,4
7	197,9±2,3	200,4±4,6	197,5±2,3	200,4±4,5
14	197,6±3,6	200,6±3,4	196,6±3,6	200,6±3,4
21	197,0±2,9	200,4±5,6	197,4±2,9	200,4±5,6
28	197,2±8,7	202,1±1,8	197,2±3,7	200,1±2,8
35	197,4±2,9	200,0±2,7	197,4±2,9	200,0±4,7
42	197,2±3,7	200,9±1,9	197,2±3,7	200,9±1,9
49	197,3±2,4	200,8±1,3	197,4±1,7	200,8±2,3
56	197,2±3,5	200,9±1,9	197,3±3,5	200,9±2,9
61	199,5±4,8	200,7±1,9	197,6±4,2	201,9±2,4

infusion doesn’t influence on behavioral responses, general state of health and frequency of eating, defecation and urination of experimental animals.

After 60 days animals were removed from the experiment by decollation under espneic anesthesia by ether vapor in accordance with rules for experimental animals [6]. Visual inspection of dead animals doesn’t identify any significant distinction from animals of control groups.

All animals had normal build and average state of nourishment. Oral mucosa and tongue were pale and clean. Teeth were preserved. Excretions from natural foramen didn’t present. Fur was shining and clean. There are no centers of alopecia, skin irritation and erosion were noticed.

Thus toxity measurement and animal observation data during 60 days let to relate C. officinalis infusion to the fourth class of low hazard substances [7].

Histological research of experimental animals internal organs in comparison with control animals internal organs were also held. Animals’ hearts, livers, kidneys and stomachs were fixed in Carnoy’s fluid, neutral formalin and flooded by paraffin for these purposes. Sections were coloured by hematoxylin and eosin by Van Gieson. In the course of researches was established:

- endothelial cells of aortha inner membrane have clear nuclea; there are no destruction of tunica media elastic fibers; myofibrills’ transverse striation is clear in all heart sections, cardiomyocites’ nuclea contain sufficient quantity of chromatin, nuclear membrane is thin. There are no centers of cytoplasm tinctorial properties violation and cardiofibrosis; slight swelling of heart’s stroma was fixed, which was associated with anesthesia (picture 1);

- trabecular liver structure on cuts from different liver lobes doesn’t detect violations. Hepatocytes’ borders are clear, cytoplasm is granular. There are no local tinctorial properties disorders of cytoplasm. Nuclea contain clear nucleola and sufficient quantity of chromatin. Nuclear membrane is thin. Liver’s sinusoids are plethoric (picture 2);

Picture 1

Histological research results

(heart’s stroma slight swelling)

Picture 2

Histological research results

(liver’s sinusoids plethora)

- capillaries of nephrons and intersticial tissue in kidneys are plethoric, cytoplasm of kidney proximal tubule epithelium is oxyphilic, cell’s borders are distinguishable, nuclea are bright and clear (picture 3);

Picture 3

Histological research results

(plethora of kidney tissue)

-integumentary epithelium of stomach mucosa is formed by mucous cylindrical cells. Epithelial lining defects were not noticed (picture 4). Principal and parietal cells of stomach glands are not changed.

Picture 4

Histological research results

(integumentary epithelium of stomach mucosa)

There were no clear differences between microscopic sections of experimental and control groups of animals were detected as result of histological research. Daily intragastric giving of C. officinalis to experimental animals of both sexes during 60 days didn’t cause irritation, inflammation and destruction of tissues. Dystrophic, destructive, local sclerotic changes in parenchymal cells and stroma of internal organs didn’t occur.

References:

1. Brock A. Brassicaceae contain nortropane alkaloids/ A. Brock, Т. Herzfeld, R. Paschke, M. Koch et all// Phytochemistry.- 2006. -Vol. 67. -Iss. 18.- Pp. 2050-2057.

2. M. G. Irapetyanz, I. P. Levshyna, L. V. Nozdracheva, N. N. Shuikyn, Correction of behavioral and physiological indexes of white rats neurosis-like state by giving the succinic acid, J. Higher nervous activity, V. 51, №3 (2001), p. 360–367.

3. RF Ministry of Health order № 267 from 19.06.2003 «Adoption of the rules of laboratory practices».

4. R. U. Habriev, O. L. Verstakova, E. V. Arzamastzev, E. A. Babayan

Guidance for experimental (preclinical) research of new pharmacological agents, (2005), p. 832.

5. State Pharmacopoeia XI, V. 2, (1990), p. 400.

6. RF Ministry of Health order № 755 from 12.08.1977 «Improvement steps of work’s organizational forms with use of experimental animals».

7. State industry standard 12.1.007-76 from 01.01.1977, System of safety standards. Harmful substances. Classification and safety requirements.